Repository logo
 

Precise optical control of gene expression in C elegans using improved genetic code expansion and Cre recombinase

Published version
Peer-reviewed

Change log

Authors

Davis, Lloyd 
Radman, Inja 
Goutou, Angeliki 
Tynan, Ailish 
Baxter, Kieran 

Abstract

Synthetic strategies for optically controlling gene expression may enable the precise spatiotemporal control of genes in any combination of cells that cannot be targeted with specific promoters. We develop an improved genetic code expansion system in Caenorhabditis elegans and use it to create a photoactivatable Cre recombinase. We laser-activate Cre in single neurons within a bilaterally symmetric pair to selectively switch on expression of a loxP-controlled optogenetic channel in the targeted neuron. We use the system to dissect, in freely moving animals, the individual contributions of the mechanosensory neurons PLML/PLMR to the C. elegans touch response circuit, revealing distinct and synergistic roles for these neurons. We thus demonstrate how genetic code expansion and optical targeting can be combined to break the symmetry of neuron pairs and dissect behavioural outputs of individual neurons that cannot be genetically targeted.

Description

Funder: Wellcome-Trust University of Edinburgh Institutional Strategic Support Fund ISS2


Funder: The Royal Society


Funder: Muir Maxwell Epilepsy Centre


Funder: Louis Jeantet Foundation


Funder: Herchel Smith Foundation


Funder: University of Edinburgh, Edinburgh Global Award and Principal's Career Development PhD studentship

Keywords

Tools and Resources, Biochemistry and Chemical Biology, Neuroscience, genetic code expansion, non canonical amino acids, optogenetics, Cre recombinase, photo-caged amino acid, C. elegans

Journal Title

eLife

Conference Name

Journal ISSN

2050-084X

Volume Title

10

Publisher

eLife Sciences Publications, Ltd
Sponsorship
H2020 European Research Council (ERC-StG-679990)
Medical Research Council (MC_U105181009)
Medical Research Council (MC_UP_A024_1008)